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Interaction between Butyrate and Tumor Necrosis Factor α in Primary Rat Colonocytes

Butyrate, a short-chain fatty acid, is utilized by the gut epithelium as energy and it improves the gut epithelial barrier. More recently, it has been associated with beneficial effects on immune and cardiovascular homeostasis. Conversely, tumor necrosis factor alpha (TNFα) is a pro-inflammatory and...

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Autores principales: Souders, Christopher L., Aristizabal-Henao, Juan J., Patuel, Sarah J., Bowden, John A., Zubcevic, Jasenka, Martyniuk, Christopher J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9953264/
https://www.ncbi.nlm.nih.gov/pubmed/36830627
http://dx.doi.org/10.3390/biom13020258
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author Souders, Christopher L.
Aristizabal-Henao, Juan J.
Patuel, Sarah J.
Bowden, John A.
Zubcevic, Jasenka
Martyniuk, Christopher J.
author_facet Souders, Christopher L.
Aristizabal-Henao, Juan J.
Patuel, Sarah J.
Bowden, John A.
Zubcevic, Jasenka
Martyniuk, Christopher J.
author_sort Souders, Christopher L.
collection PubMed
description Butyrate, a short-chain fatty acid, is utilized by the gut epithelium as energy and it improves the gut epithelial barrier. More recently, it has been associated with beneficial effects on immune and cardiovascular homeostasis. Conversely, tumor necrosis factor alpha (TNFα) is a pro-inflammatory and pro-hypertensive cytokine. While butyrate and TNFα are both linked with hypertension, studies have not yet addressed their interaction in the colon. Here, we investigated the capacity of butyrate to modulate a host of effects of TNFα in primary rodent colonic cells in vitro. We measured ATP levels, cell viability, mitochondrial membrane potential (MMP), reactive oxygen species (ROS), mitochondrial oxidative phosphorylation, and glycolytic activity in colonocytes following exposure to either butyrate or TNFα, or both. To address the potential mechanisms, transcripts related to oxidative stress, cell fate, and cell metabolism (Pdk1, Pdk2, Pdk4, Spr, Slc16a1, Slc16a3, Ppargc1a, Cs, Lgr5, Casp3, Tnfr2, Bax, Bcl2, Sod1, Sod2, and Cat) were measured, and untargeted liquid chromatography–tandem mass spectrometry (LC-MS/MS) was employed to profile the metabolic responses of colonocytes following exposure to butyrate and TNFα. We found that both butyrate and TNFα lowered cellular ATP levels towards a quiescent cell energy phenotype, characterized by decreased oxygen consumption and extracellular acidification. Co-treatment with butyrate ameliorated TNFα-induced cytotoxicity and the reduction in cell viability. Butyrate also opposed the TNFα-mediated decrease in MMP and mitochondrial-to-intracellular calcium ratios, suggesting that butyrate may protect colonocytes against TNFα-induced cytotoxicity by decreasing mitochondrial calcium flux. The relative expression levels of pyruvate dehydrogenase kinase 4 (Pdk4) were increased via co-treatment of butyrate and TNFα, suggesting the synergistic inhibition of glycolysis. TNFα alone reduced the expression of monocarboxylate transporters slc16a1 and slc16a3, suggesting effects of TNFα on butyrate uptake into colonocytes. Of the 185 metabolites that were detected with LC-MS, the TNFα-induced increase in biopterin produced the only significant change, suggesting an alteration in mitochondrial biogenesis in colonocytes. Considering the reports of elevated colonic TNFα and reduced butyrate metabolism in many conditions, including in hypertension, the present work sheds light on cellular interactions between TNFα and butyrate in colonocytes that may be important in understanding conditions of the colon.
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spelling pubmed-99532642023-02-25 Interaction between Butyrate and Tumor Necrosis Factor α in Primary Rat Colonocytes Souders, Christopher L. Aristizabal-Henao, Juan J. Patuel, Sarah J. Bowden, John A. Zubcevic, Jasenka Martyniuk, Christopher J. Biomolecules Article Butyrate, a short-chain fatty acid, is utilized by the gut epithelium as energy and it improves the gut epithelial barrier. More recently, it has been associated with beneficial effects on immune and cardiovascular homeostasis. Conversely, tumor necrosis factor alpha (TNFα) is a pro-inflammatory and pro-hypertensive cytokine. While butyrate and TNFα are both linked with hypertension, studies have not yet addressed their interaction in the colon. Here, we investigated the capacity of butyrate to modulate a host of effects of TNFα in primary rodent colonic cells in vitro. We measured ATP levels, cell viability, mitochondrial membrane potential (MMP), reactive oxygen species (ROS), mitochondrial oxidative phosphorylation, and glycolytic activity in colonocytes following exposure to either butyrate or TNFα, or both. To address the potential mechanisms, transcripts related to oxidative stress, cell fate, and cell metabolism (Pdk1, Pdk2, Pdk4, Spr, Slc16a1, Slc16a3, Ppargc1a, Cs, Lgr5, Casp3, Tnfr2, Bax, Bcl2, Sod1, Sod2, and Cat) were measured, and untargeted liquid chromatography–tandem mass spectrometry (LC-MS/MS) was employed to profile the metabolic responses of colonocytes following exposure to butyrate and TNFα. We found that both butyrate and TNFα lowered cellular ATP levels towards a quiescent cell energy phenotype, characterized by decreased oxygen consumption and extracellular acidification. Co-treatment with butyrate ameliorated TNFα-induced cytotoxicity and the reduction in cell viability. Butyrate also opposed the TNFα-mediated decrease in MMP and mitochondrial-to-intracellular calcium ratios, suggesting that butyrate may protect colonocytes against TNFα-induced cytotoxicity by decreasing mitochondrial calcium flux. The relative expression levels of pyruvate dehydrogenase kinase 4 (Pdk4) were increased via co-treatment of butyrate and TNFα, suggesting the synergistic inhibition of glycolysis. TNFα alone reduced the expression of monocarboxylate transporters slc16a1 and slc16a3, suggesting effects of TNFα on butyrate uptake into colonocytes. Of the 185 metabolites that were detected with LC-MS, the TNFα-induced increase in biopterin produced the only significant change, suggesting an alteration in mitochondrial biogenesis in colonocytes. Considering the reports of elevated colonic TNFα and reduced butyrate metabolism in many conditions, including in hypertension, the present work sheds light on cellular interactions between TNFα and butyrate in colonocytes that may be important in understanding conditions of the colon. MDPI 2023-01-30 /pmc/articles/PMC9953264/ /pubmed/36830627 http://dx.doi.org/10.3390/biom13020258 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Souders, Christopher L.
Aristizabal-Henao, Juan J.
Patuel, Sarah J.
Bowden, John A.
Zubcevic, Jasenka
Martyniuk, Christopher J.
Interaction between Butyrate and Tumor Necrosis Factor α in Primary Rat Colonocytes
title Interaction between Butyrate and Tumor Necrosis Factor α in Primary Rat Colonocytes
title_full Interaction between Butyrate and Tumor Necrosis Factor α in Primary Rat Colonocytes
title_fullStr Interaction between Butyrate and Tumor Necrosis Factor α in Primary Rat Colonocytes
title_full_unstemmed Interaction between Butyrate and Tumor Necrosis Factor α in Primary Rat Colonocytes
title_short Interaction between Butyrate and Tumor Necrosis Factor α in Primary Rat Colonocytes
title_sort interaction between butyrate and tumor necrosis factor α in primary rat colonocytes
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9953264/
https://www.ncbi.nlm.nih.gov/pubmed/36830627
http://dx.doi.org/10.3390/biom13020258
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